BD HorizonTM Fixable Viability Stain 780

Description

Figure 1. Fluorescent staining of Jurkat cells with BD Horizon Fixable Viability Stain 780. Human Jurkat cells were treated 16 hr with 0.025 % DMSO Top Plot or 5 muM camptothecin Bottom Plot and stained with BD Horizon Fixable Viability Stain 780 Cat. No. 565388 . Cells were either not fixed solid line histograms , or fixed in BD Cytofix Fixation Buffer Cat. No. 554655 and permeabilized in Perm/Wash Buffer I Cat. No. 557885 dashed line histograms . Histograms were derived from gated events with the light scattering characteristics of Jurkat cells. Flow cytometry was performed using a BD LSRII Cell Analyzer System. Figure 2. Analysis of proliferating mouse splenocytes for surface and intracellular markers. BALB/c splenocytes were stained 10 min, 37C with BD Horizon Violet Proliferation Dye 450 Cat. No. 562158 , washed twice, and then cultured 3 days with Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057 and Hamster Anti-Mouse CD28 Cat. No. 553294 antibodies. The cells were restimulated 4 hr with PMA, Ionomycin, and BD GolgiStop Protein Transport Inhibitor Monensin Cat. No. 554724 . Cells were harvested, stained with BD Horizon Fixable Viability Stain 780, fixed and permeabilized using a BD Cytofix/Cytoperm Fixation/Permeabilization Solution Kit Cat. No. 554714 , and then stained with BD Horizon BUV395 Anti-Mouse CD4 Cat. No. 563790 and FITC Anti-Mouse IL-2 Cat. No. 554427 antibodies. Two-color dot plots showing VPD450 fluorescence versus CD4 expression Middle Plots or IL-2 expression Right Plots were derived from either total cells Top; FVS780 Dull and Bright Cells or previously viable cells Bottom; FVS780 Dull Cells with the light scatter characteristics of intact cells. CD4+ and IL-2+ cell gates were based on an FMO or unstimulated cell control, respectively. Flow cytometry was performed on a BD LSRFortessa Cell Analyzer system. Figure 1. Fluorescent staining of Jurkat cells with BD Horizon Fixable Viability Stain 780. Human Jurkat cells were treated 16 hr with 0.025 % DMSO Top Plot or 5 muM camptothecin Bottom Plot and stained with BD Horizon Fixable Viability Stain 780 Cat. No. 565388 . Cells were either not fixed solid line histograms , or fixed in BD Cytofix Fixation Buffer Cat. No. 554655 and permeabilized in Perm/Wash Buffer I Cat. No. 557885 dashed line histograms . Histograms were derived from gated events with the light scattering characteristics of Jurkat cells. Flow cytometry was performed using a BD LSRII Cell Analyzer System. Figure 2. Analysis of proliferating mouse splenocytes for surface and intracellular markers. BALB/c splenocytes were stained 10 min, 37C with BD Horizon Violet Proliferation Dye 450 Cat. No. 562158 , washed twice, and then cultured 3 days with Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057 and Hamster Anti-Mouse CD28 Cat. No. 553294 antibodies. The cells were restimulated 4 hr with PMA, Ionomycin, and BD GolgiStop Protein Transport Inhibitor Monensin Cat. No. 554724 . Cells were harvested, stained with BD Horizon Fixable Viability Stain 780, fixed and permeabilized using a BD Cytofix/Cytoperm Fixation/Permeabilization Solution Kit Cat. No. 554714 , and then stained with BD Horizon BUV395 Anti-Mouse CD4 Cat. No. 563790 and FITC Anti-Mouse IL-2 Cat. No. 554427 antibodies. Two-color dot plots showing VPD450 fluorescence versus CD4 expression Middle Plots or IL-2 expression Right Plots were derived from either total cells Top; FVS780 Dull and Bright Cells or previously viable cells Bottom; FVS780 Dull Cells with the light scatter characteristics of intact cells. CD4+ and IL-2+ cell gates were based on an FMO or unstimulated cell control, respectively. Flow cytometry was performed on a BD LSRFortessa Cell Analyzer system. Show More Figure 1. Fluorescent staining of Jurkat cells with BD Horizon Fixable Viability Stain 780. Human Jurkat cells were treated 16 hr with 0.025 % DMSO Top Plot or 5 muM camptothecin Bottom Plot and stained with BD Horizon Fixable Viability Stain 780 Cat. No. 565388 . Cells were either not fixed solid line histograms , or fixed in BD Cytofix Fixation Buffer Cat. No. 554655 and permeabilized in Perm/Wash Buffer I Cat. No. 557885 dashed line histograms . Histograms were derived from gated events with the light scattering characteristics of Jurkat cells. Flow cytometry was performed using a BD LSRII Cell Analyzer System. Figure 2. Analysis of proliferating mouse splenocytes for surface and intracellular markers. BALB/c splenocytes were stained 10 min, 37C with BD Horizon Violet Proliferation Dye 450 Cat. No. 562158 , washed twice, and then cultured 3 days with Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057 and Hamster Anti-Mouse CD28 Cat. No. 553294 antibodies. The cells were restimulated 4 hr with PMA, Ionomycin, and BD GolgiStop Protein Transport Inhibitor Monensin Cat. No. 554724 . Cells were harvested, stained with BD Horizon Fixable Viability Stain 780, fixed and permeabilized using a BD Cytofix/Cytoperm Fixation/Permeabilization Solution Kit Cat. No. 554714 , and then stained with BD Horizon BUV395 Anti-Mouse CD4 Cat. No. 563790 and FITC Anti-Mouse IL-2 Cat. No. 554427 antibodies. Two-color dot plots showing VPD450 fluorescence versus CD4 expression Middle Plots or IL-2 expression Right Plots were derived from either total cells Top; FVS780 Dull and Bright Cells or previously viable cells Bottom; FVS780 Dull Cells with the light scatter characteristics of intact cells. CD4+ and IL-2+ cell gates were based on an FMO or unstimulated cell control, respectively. Flow cytometry was performed on a BD LSRFortessa Cell Analyzer system.